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culture expansion
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  扩大培养
     Methods We select 3 sites in the complete coden sequence region of β-catenin gene as the RNAi targets, ligated the annealed double pre-DNA strands into the retroviral vectors pSUPER-retro and transfected them into the packaging cells PA317, and then collected supernatant with retrovirus to infect DU145. After selection by puromycin and culture expansion, the stable cell clones were attained.
     方法:在βcatenin基因编码区选择3个RNAi作用的靶位点,体外合成前体DNA链,复性后连入逆转录病毒载体pSUPERretro,转染包装细胞PA317,收集含病毒颗粒的培养上清感染DU145细胞,经嘌呤霉素筛选并扩大培养后得到克隆,继续培养2个月形成稳定克隆。
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  “culture expansion”译为未确定词的双语例句
     Cross-culture Theory for the Study of TCL Mobile Phone Brand Culture Expansion
     跨文化理论导入TCL手机品牌文化推广研究
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     American Mass Culture Expansion and Development of The Third World
     美国大众文化传播与第三世界国家发展
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     Methods By using a series of techniques, i.e., stem/progenitor cells culture, expansion in vitro and transplantation in animal model, the hematopoietic activity of fractionated cells in discontinuous Ficoll gradient was evaluated.
     方法 利用造血干 /祖细胞培养、体外扩增、移植动物模型等技术 ,研究不同比密ficoll分离液分离的脐血造血干 /祖细胞的造血活性。
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     Objective To establish a method for isolation and culture of bone marrow mesenchymal stem cells(MSC) from Lewis rat in vitro and to identify characteristic of the cells after culture expansion.
     目的建立从Lewis大鼠骨髓中分离、培养间充质干细胞的方法,并对分离、培养的间充质干细胞进行鉴定。
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     Objective:1 The aim of this study is to establish the method for isolation and in vitro culture expansion of adult human bone marrow-derived mesenchymal stem cells (MSCs) and to investigate their biological characteristics.
     1 建立一种从成人骨髓中分离和扩增间充质干细胞(MSC)的方法,并从细胞形态、抗原表达、超微结构、体外分化能力、增殖方式和细胞周期等方面研究其生物学特性,并对其加以鉴定。
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  相似匹配句对
     CULTURE
     论文化
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     CULTURE
     文化·流行
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     On the Expansion and Influence of Laohushan Culture
     老虎山文化的扩张与对外影响
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     The Expansion of Western Culture and Cultural "Westernization"
     西方文化扩张与文化的“泛西方化”
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     On the Earth expansion
     地球膨胀说提出、发展及其主要事实依据
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  culture expansion
Expression of Tbx3 was downregulated during culture expansion.
      
In this study, we demonstrate that bioreactors generate cells, which upon replating into secondary bioreactors, lead to continued cell, CFU-GM, and LTC-IC8 (measured after 8 weeks of secondary culture) expansion.
      
The CD9 expression in hASCs was down-regulated during culture expansion.
      
In this study, we describe a simple, high-yield procedure, requiring minimal culture expansion, for the isolation of mesenchymal progenitor cells from human trabecular bone.
      
Thus, the RQPR motif occurred in a small population of naturally occurring hMPV and was not necessarily induced during tissue culture expansion.
      
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Objective To study and establish the method for isolation,culture,proliferation and identification of canine bone marrow mesenchymal stem cells (MSCs).MethodsTen ml canine bone marrow aspirate,taken from the iliac crest of normal canines,were diluted 1∶1 with Dullbecco's Modified Eagle Medium (DMEM).The washed cells were resuspended in DMEM to a final volume of 10 ml and layered over an equal volume of 1.063 g/ml Percoll gradient solution.After centrifugation at 600 g for 30 minutes,the mononuclear cells(MNCs)were...

Objective To study and establish the method for isolation,culture,proliferation and identification of canine bone marrow mesenchymal stem cells (MSCs).MethodsTen ml canine bone marrow aspirate,taken from the iliac crest of normal canines,were diluted 1∶1 with Dullbecco's Modified Eagle Medium (DMEM).The washed cells were resuspended in DMEM to a final volume of 10 ml and layered over an equal volume of 1.063 g/ml Percoll gradient solution.After centrifugation at 600 g for 30 minutes,the mononuclear cells(MNCs)were recovered from the gradient interface and washed with PBS.Percoll fractionated MNCs were suspended in DMEM containing 1 g/L of glucose supplemented with 10% fetal bovine serum,100 U/ml penicillin,100 μg/ml streptomycin,and 25 μg/ml amphotericin B.All cells were plated in 10 ml of medium in a culture dishes.The cultures were maintained at 37℃ in air of 5% CO 2,with an initial medium change at 24 hours after initial plating and then medium change every 3 or 4 days.The MSCs were identified by special cellular surface antigens and pluripotent committing differentiation potential.Results The methodology for isolation and expansion of MSCs was established.The mononuclear cells from canine bone marrow were separated on Percoll gradient sedimentation solution at 600 g for 30 minutes in small percentage (about 0.01~0.001%).The MSCs were set very good in culture of LG-DMEM supplemented with 10% selected fetal bovine serum.MSCs attached and grew as fibroblastic cells at 24 hours after initial plating,about 10 -6 out of mononuclear cells.The hematopoietic stem cells that did not attach to the dish were washed from the culturewith each medium change.MSCs grew rapidly,developed into visible symmetric colonies at about 3 days and reached confluence at 7 to 10 days.While the cells were permitted to proliferate to confluence,MSCs were spindle-shaped morphology,small and arranged like pectinate.As many as(1.1~1.5)×10 7 cells were generated by passage 3 from a 10 ml marrow aspirate.The MSCs did not differentiate spontaneously during culture expansion and maintained a normal mesenchymal stem cells biological features.These expanded attached mesenchymal stem cells were positive for SH2,and negative for CD45 surface antigen.The mesenchymal stem cells have shown the ability to give rise to a varity of differentiated cell types such as cardiomyocytes,adipocytes and osteocytes.Conclusions The mesenchymal stem cells from canine bone marrow could be generated,cultured and expanded in vitro.These cells displayed a stable phenotype,biological and cellular features.The cultivation and selective differentiation of MSCs should provide a new cell source for stem cells transplantation,and the potential of new therapeutic approaches for the restoration of damaged or diseased tissue.

目的 建立犬骨髓间叶干细胞的体外分离、培养、扩增与鉴定方法 ,了解其生物学特性 ,为心肌梗死、心力衰竭及缓慢性心律失常的干细胞移植提供细胞材料。方法 抽取犬骨髓液 10ml,以DMEM 1∶1稀释 ,用 1 0 6 3g/mlpercoll密度分离液 ,以 6 0 0 g离心力、30min分离骨髓单个核细胞 ;以LG DMEM及 10 %FBS添加 10 0U/ml青霉素 ,10 0 μg/ml链霉素 ,2 5 μg/ml两性霉素B培养骨髓干细胞 ;利用细胞贴壁筛选法分离、培养、扩增骨髓间叶干细胞 ;应用干细胞表面标志蛋白检测 ,诱导剂诱导分化间叶组织细胞等方法进行犬骨髓间叶干细胞的鉴定。结果 分离出的犬骨髓单个核细胞约占细胞总数的 10 -4~ 10 -6,在LG DMEM与选择性血清培养基上生长良好。犬骨髓间叶干细胞孵育 2 4h即可见细胞贴壁生长 ,贴壁细胞约占接种单个核细胞总数的 10 -6。犬骨髓间叶干细胞增殖分裂迅速 ,38~ 4 8h内增殖多个细胞 ,约 72h即可增殖形成大的细胞克隆 ,7~ 10d即可布满瓶底。细胞融合时类似成纤维细胞 ,呈纺锤状 ,细胞小而密集 ,螺旋梳状排列。连续培育 1...

目的 建立犬骨髓间叶干细胞的体外分离、培养、扩增与鉴定方法 ,了解其生物学特性 ,为心肌梗死、心力衰竭及缓慢性心律失常的干细胞移植提供细胞材料。方法 抽取犬骨髓液 10ml,以DMEM 1∶1稀释 ,用 1 0 6 3g/mlpercoll密度分离液 ,以 6 0 0 g离心力、30min分离骨髓单个核细胞 ;以LG DMEM及 10 %FBS添加 10 0U/ml青霉素 ,10 0 μg/ml链霉素 ,2 5 μg/ml两性霉素B培养骨髓干细胞 ;利用细胞贴壁筛选法分离、培养、扩增骨髓间叶干细胞 ;应用干细胞表面标志蛋白检测 ,诱导剂诱导分化间叶组织细胞等方法进行犬骨髓间叶干细胞的鉴定。结果 分离出的犬骨髓单个核细胞约占细胞总数的 10 -4~ 10 -6,在LG DMEM与选择性血清培养基上生长良好。犬骨髓间叶干细胞孵育 2 4h即可见细胞贴壁生长 ,贴壁细胞约占接种单个核细胞总数的 10 -6。犬骨髓间叶干细胞增殖分裂迅速 ,38~ 4 8h内增殖多个细胞 ,约 72h即可增殖形成大的细胞克隆 ,7~ 10d即可布满瓶底。细胞融合时类似成纤维细胞 ,呈纺锤状 ,细胞小而密集 ,螺旋梳状排列。连续培育 10代以上 ,未见细胞形态、增殖特性发生改变。未见骨髓间叶干细胞自发分化其它类型细胞 ,仍维持原代培养细胞的增殖特性。犬骨髓间叶干细胞的表面标志SH2阳性 ,CD4 5则阴性。在化学诱导

Objective To set up a prostate cancer cell line in which β-catenin expression is stably suppressed and to investigate the role of Wnt/β-catenin signaling pathway in prostate tumorgenesis. Methods We select 3 sites in the complete coden sequence region of β-catenin gene as the RNAi targets, ligated the annealed double pre-DNA strands into the retroviral vectors pSUPER-retro and transfected them into the packaging cells PA317, and then collected supernatant with retrovirus to infect DU145. After selection by puromycin...

Objective To set up a prostate cancer cell line in which β-catenin expression is stably suppressed and to investigate the role of Wnt/β-catenin signaling pathway in prostate tumorgenesis. Methods We select 3 sites in the complete coden sequence region of β-catenin gene as the RNAi targets, ligated the annealed double pre-DNA strands into the retroviral vectors pSUPER-retro and transfected them into the packaging cells PA317, and then collected supernatant with retrovirus to infect DU145. After selection by puromycin and culture expansion, the stable cell clones were attained. Expression of the 2 target genes of Wnt/β-catenin signaling pathway cyclinD1 and c-myc, was detected in the β-catenin RNAi cells by Western blot. The effect of suppressing β-catenin by RNAi on cell proliferation was quantified by methylthiazoletetrazolium (MTT) assay. Results Western blotting and RT-PCR showed that the expression level of β-catenin in the 2 stable cell clones apparently decreased. CyclinD1 and c-myc expression decreased in the β-catenin RNAi cells. MTT showed that the cell number of β-catenin expression suppression cell clones decreased significantly (P<0.05), suggesting the cell proliferation was prevented. Conclusion The β-catenin gene stable suppression cell line was successfully established.

目的:建立稳定抑制βcatenin基因表达的前列腺癌细胞株,为探讨Wnt/βcatenin信号在前列腺癌发生中的作用提供合适的细胞模型。方法:在βcatenin基因编码区选择3个RNAi作用的靶位点,体外合成前体DNA链,复性后连入逆转录病毒载体pSUPERretro,转染包装细胞PA317,收集含病毒颗粒的培养上清感染DU145细胞,经嘌呤霉素筛选并扩大培养后得到克隆,继续培养2个月形成稳定克隆。免疫印迹检测癌细胞内βcatenin及受其调控的靶基因cmyc和cyclinD1的表达;四甲基偶氮唑盐(MTT)比色法检测βcatenin基因表达抑制的细胞和对照组细胞的增殖。结果:免疫印迹和RTPCR结果显示,筛选得到的2个克隆中细胞内βcatenin表达水平下降;且克隆内cmyc,cyclinD1基因的表达下调;MTT检测显示,βcatenin表达抑制的细胞克隆吸光度值下降,这表明克隆内细胞数量减少,细胞增殖受到了抑制。结论:成功建立稳定抑制βcatenin基因表达的前列腺癌细胞株。

Objective The present study was aimed to investigate the appropriate and reliable methods for isolation, culture-expansion of canine bone marrow mesenchymal stem cells (MSCs). Methods The bone marrow mononuclear cells were isolated from 15 -20 ml canine bone marrow aspirates, by using density gradient centrifugation with 1.068 g/ml or 1.073 g/ml separating mediums. All cells were cultured at 37℃ under5% CO2 atmosphere in DMEM supplemented with 15% fetal bovine serum, 100 U/ml penicillin, 100 μg/ml...

Objective The present study was aimed to investigate the appropriate and reliable methods for isolation, culture-expansion of canine bone marrow mesenchymal stem cells (MSCs). Methods The bone marrow mononuclear cells were isolated from 15 -20 ml canine bone marrow aspirates, by using density gradient centrifugation with 1.068 g/ml or 1.073 g/ml separating mediums. All cells were cultured at 37℃ under5% CO2 atmosphere in DMEM supplemented with 15% fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, and 25 μg/ml amphotericin B. The medium was replaced every 2 or 3 days. The MSCs were identified by special cellular surface antigens and pluripotent differentiation potential. Results With the density gradient centrifugation method, the mononuclear cells were successfully isolated, and the Nycoprep 1. 068 group could obtain more purified monocytes. The adherent cells proliferated quickly at a doubling time of 24 hr. When the cells were confluent, the morphology of adherent cells was spindle-shaped, small and arranged like pectinate. The characteristics of proliferation and morphology did not change after 8 passages, and there was no evidence of spontaneous differentiation. The expanded adherent cells were positive for SH2, and negative for CD45 surface antigen. Under the induction of chemicals, the MSCs differentiated into cardiomyocytes, adipocytes and osteocytes. Conclusions The mesenchymal stem cells from canine bone marrow could be isolated, cultured and expanded in vitro through the density gradient centrifugation with Nycoprep?1.068 and adherent cell cultivation. These cells showed the characteristics of mesenchymal stem cells, with self-replenishing ability and pluripotent differentiation potential.

目的探讨更为可靠、纯净度高的犬骨髓间充质干细胞分离、培养方法。方法将犬骨髓抽取液分别以1.068g/ml及1.073g/ml分离液进行密度梯度离心,采用贴壁培养法获得骨髓间充质干细胞(MSCs);以LG-DMEM加15%FBS培养;应用干细胞表面标志蛋白,多向诱导分化等方法进行细胞鉴定。结果采用1.068g/ml分离液可获得纯度较高的单核细胞,接种细胞生长良好,孵育24h即可见细胞贴壁生长,平均倍增周期为1d,细胞呈纺锤状,螺旋梳状排列。连续培育8代以上,未见细胞形态、增殖特性改变。MSCs表面标志SH2阳性,CD45阴性,可定向分化为心肌细胞、成骨细胞及脂肪细胞。结论采用1.068s/ml分离液能够较好地进行犬MSCs的体外分离、培养与扩增,分离得到的细胞具备MSCs的特性。

 
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